Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite bovino produzido no sistema orgânico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation of Staphylococcus epidermidis from inflamed upper respiratory tract of an orange-spined hairy dwarf porcupine (Sphiggurus villosus)

The orange-spined hairy dwarf porcupine (Sphiggurus villosus) is a rodent species common in most parts of South America, and little is known about the pathologies that can afflict it. A specimen was delivered at the Wildlife Research and Medical Center (CEMPAS), School of Veterinary Medicine and Animal Husbandry, UNESP, Botucatu, SP, Brazil. The animal showed intense apathy, with purulent secretion in the nasal cavity and fracture of the lumbar spine. Due to the unfavorable prognosis, the porcupine was euthanized and microbiological culture of nasal discharge showed Staphylococcus epidermidis. The antimicrobial resistance test revealed sensitivity to all tested antimicrobials (ampicillin, oxacillin, tetracycline, penicillin G, neomycin, cephalexin, gentamicin, enrofloxacin, ciprofloxacin, cotrimoxazol, cefoxitin and cephalothin). This bacterium is part of the nasal flora of humans and other animals, and may cause infection under certain conditions. In the present study, the infection and colonization by S. epidermidis was the probable cause of the inflammatory process. The sensitivity to all tested antimicrobials suggests that this strain has not been previously exposed to such drugs.

Predictive factors of outcome following staphylococcal peritonitis in continuous ambulatory peritoneal dialysis

Background and aims: Staphylococcus epidermidis and other coagulase-negative staphylococci (CoNS) are the most common agents of continuous ambulatory peritoneal dialysis (CAPD) peritonitis. Episodes caused by Staphylococcus aureus evolve with a high method failure rate while CoNS peritonitis is generally benign. The purpose of this study was to compare episodes of peritonitis caused by CoNS species and S. aureus to evaluate the microbiological and host factors that affect outcome. Material and methods: Microbiological and clinical data were retrospectively studied from 86 new episodes of peritonitis caused by staphylococci species between January 1996 and December 2000 in a university dialysis center. The influence of microbiological and host factors (age, sex, diabetes, use of vancomycin, exchange system and treatment time on CAPD) was analyzed by logistic regression model. The clinical outcome was classified into two results (resolution and non-resolution). Results: the odds of peritonitis resolution were not influenced by host factors. Oxacillin susceptibility was present in 30 of 35 S. aureus lineages and 22 of 51 CoNS (p = 0.001). There were 32 of 52 (61.5%) episodes caused by oxacillin-susceptible and 20 of 34 (58.8%) by oxacillin-resistant lineages resolved (p = 0.9713). of the 35 cases caused by S. aureus, 17 (48.6%) resolved and among 51 CoNS episodes 40 (78.4%) resolved. Resolution odds were 7.1 times higher for S. epidermidis than S. aureus (p = 0.0278), while other CoNS had 7.6 times higher odds resolution than S. epidermidis cases (p = 0.052). Episodes caused by S. haemolyticus had similar resolution odds to S. epidermidis (p = 0.859). Conclusions: S. aureus etiology is an independent factor associated with peritonitis non-resolution in CAPD, while S. epidermidis and S. haemolyticus have a lower resolution rate than other CoNS. Possibly the aggressive nature of these agents, particularly S. aureus, can be explained by their recognized pathogenic factors, more than antibiotic resistance.

Evalution of methods for the detection of MRSA in patients of Botucatu Medical School Hospital, São Paulo, Brazil

Oxacillin is the main drug of choice for the treatment of S. aureus infections. However, S. aureus resistance to oxacillin has become a major problem in the recent decades. The study aimed assess the rates of oxacillin resistance in S. aureus samples obtained at the Botucatu Medical School Hospital, UNESP, and to compare phenotypic techniques for the detection of MRSA against the gold standard method (mecA gene detection) in these samples. A total of 102 samples, previously isolated between 2002 and 2006, and kept at the Culture Collection of the Department of Microbiology and Immunology, in the Botucatu Biosciences Institute, UNESP, were included. Oxacillin resistance was assessed by oxacillin and cefoxitin disk diffusion and agar dilution tests, screening tests using Mueller-Hinton agar with 6 mu g/mL of oxacillin and 4% NaCl, E-test, and mecA gene detection. of the samples analyzed, 46 (45.1%) were mecA-positive. Oxacillin disk sensitivity and specificity were 86.9% and 91.1%, respectively. Cefoxitin disk sensitivity and specificity were respectively 91.3% and 91.1%. The screening test with the cefoxitin disk showed almost the same level of sensitivity (91.3%) and specificity (91.1%). With E-test strips, sensitivity was higher (97.8%) and specificity was comparable to that found with the other methods (91.1%). Ninety-three percent of the samples produced beta-lactamase and five of them were mecA-negative. There was a gradual increase in the number of oxacillin-resistant S. aureus samples between 2002 and 2004. However, from 2004 to 2006, the number of resistant samples dropped from 55% of MRSA in 2004, to 45% in 2005 and 34.6% in 2006. The data obtained reveal that, among phenotypic methods, the E-test yielded the best results, with higher sensitivity levels when compared to the other methods. The decreased resistance rate observed over the most recent years may be explained by the rational use of antimicrobial agents associated with good practices in the control of hospital infection, or may be related to the diminished use of oxacillin as a treatment option.

Identification and antimicrobial susceptibility of Staphylococcus from home-treated peritoneal dialysis patients

Staphylococcus aureus is the main agent of infections during peritoneal dialysis (PD). The presence of S. aureus in the nasal cavity has been extensively studied and suggested as a risk factor of dialysis-related infections, whereas coagulase-negative Staphylococcus (CNS) species are frequently considered part of the normal human microbiota. The aim of this study was to identify Staphylococcus in the nasal cavity, pericatheter skin and peritoneal effluent from PD patients, as well as to evaluate the antimicrobial activity evolution in vitro. Thirty-two chronic PD patients were observed during 12 months and had nasal and pericatheter skin samples collected for culture. When peritonitis was detected, samples were also collected from the peritoneal effluent for culture. The activity of several antimicrobial drugs (penicillin G, oxacillin, cephalothin, ofloxacin, netilmicin and vancomycin) against different Staphylococcus species was measured by using the agar drug diffusion assay (Kirby-Bauer method). Staphylococcus was separated into S. aureus, S. epidermidis and other CNS species in order to determine the in vitro resistance level. S. epidermidis resistance to oxacillin progressively increased during the study period (p < 0.05). Resistance to ofloxacin was inexpressive, whereas resistance to netilmicin and vancomycin was not detected. of the oxacillin-resistant species (n = 74), 83% were S. epidermidis, 13% other CNS and 4% S. aureus (p < 0.05). Regarding multidrug resistant strains (n = 45), 82% were S. epidermidis, 13% other CNS, and 5% S. aureus (p < 0.05). This study shows the relevance of resistance to oxacillin and CNS multi-drug resistance, particularly concerning S. epidermidis, in PD patients.

Phenotype characterization of Staphylococcus species strains isolated from buffalo (Bubalus bubalis) milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da colonização nasal por Staphylococcus spp. resistenteà oxacilina em alunos de enfermagem

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dimetilsulfóxido - DMSO no teste de sensibilidade microbiana in vitro em cepas de Rhodococcus equi isoladas de afecções pulmonares em potros

Comparou-se a sensibilidade microbiana in vitro de isolados de Rhodococcus equi pelo teste padrão de difusão com discos, com o modificado, pela adição de 5% de dimetilsulfóxido-DMSO. Observou-se aumento da sensibilidade do R. equi no teste com DMSO, frente a aminoglicosídeos (canamicina, amicacina, estreptomicina) e ao cloranfenicol, enquanto para a eritromicina e derivados ß-lactâmicos (penicilina G, cefalosporinas, amoxicilina, oxacilina), constatou-se redução da sensibilidade do agente.

Semi-selective culture medium for Curtobacterium flaccumfaciens pv. flaccumfaciens isolation from bean seeds

Curtobacterium wilt has become an important disease of beans in several localities in the country. Its causal agent, Curtobacterium flaccumfacciens pv. flaccumfaciens (Cff), survives and is disseminated through seeds. To date, few studies have been conducted with the objective of developing an effective and low-cost culture medium to isolate this bacterium from bean seeds, for health analysis purposes. Usually, the culture media employed for coryneform bacteria isolation contain specific carbon sources and antimicrobial products not available in the Brazilian market. A culture medium known as MSCFF was developed (peptone - 5 g, meat extract - 3 g, sucrose - 5 g, agar 15 g, skim milk powder* - 5 g. Congo red* - 0.05 g-, chlorothalonil* - 0.01 g, thiophanate methyl* - 0.01 g, nalidixic acid* - 0.01 g, nitrofurantoin* - 0.01 g. oxacillin* 0.001 g, sodium azide* - 0.001 g and distilled water q.s. 1L; *added after autoclaving the basal medium), which has the ability to inhibit growth of a large amount of saprophytic bacteria, but with low supressivity to Cff isolates. The MSCFF medium was highly effective for Cff isolation from naturally infected bean seeds and could be used for routine detection of this bacterium in bean seeds.

Evaluation of the in vitro activity of cefepime compared to other broad-spectrum cephalosporins against clinical isolates from eighteen Brazilian hospitals by using the Etest

The in vitro activity of cefepime was compared to that of ceftazidime, ceftriaxone, and cefotaxime in a multicenter study involving 10 clinical microbiology laboratories and clinical isolates from 18 Brazilian hospitals from 7 cities (4 states). A total of 982 isolates consecutively collected between December 1995 and March 1996 were susceptibility tested by using Etest and following the NCCLS procedures for agar diffusion tests. The cefepime spectrum was broader than that of the other broad-spectrum cephalosporins against both Gram-negative rods and Gram-positive cocci. Cefepime tons particularly move active against Enterobacter sp. (MIC90, 2 mu g/ml), Serratia sp. (MIC90, 2 mu g/ml) and oxacillin-susceptible Staphylococcus aureus (MIC90, 3 mu g/ml). Against Pseudomonas aeruginosa, cefepime (MIC90 16 mu g/ml) was slightly more active than ceftazidime (MIC90 32 mu g/ml) and 8- to 16-fold more active than ceftriaxone or cefotaxime (MIC90 >256 mu g/ml). Our results show that nosocomial bacteria, especially Gram-negative rods, have a high rate of cephalosporin resistance in Brazil. However, part of these resistant bacteria remains susceptible to cefepime. The Etest was shown to be an excellent method for multicenter studies of the in vitro evaluation of new antimicrobial agents. (C) 1997 Elsevier B.V.

First report of vancomycin-resistant staphylococci isolated from healthy carriers in Brazil

Reduced susceptibility or resistance to vancomycin has been reported among clinical isolates of staphylococci in previous studies. In the present study we report on the isolation of four vancomycin-resistant staphylococcal strains from healthy carriers inside and outside the hospital environment. These carriers did not receive treatment with any antibiotic. All coagulase-negative staphylococcal strains showed variable levels of resistance to several antimicrobial agents, including oxacillin, and unstable resistance to vancomycin, with decreased vancomycin MICs (<4 mg/liter) after 10 days of passage in a nonselective medium. However, exposure of these revertants to vancomycin selected staphylococcal strains resistant to vancomycin at very high frequencies (10(-2) and 10(-3)). The vancomycin resistance in these staphylococcal strains was not mediated by the van gene. The cell wall of the staphylococcal strains studied became thickest after culture in medium containing vancomycin, and the differences in cell wall thickness were statistically significant (P < 0.001). Thus, the thickening of the cell wall in these staphylococcal strains may be an important contributor to vancomycin resistance.

Ocorrência de mastite bovina causada por Staphylococcus sp., Streptococcus sp. E Candida sp. em uma propriedade rural no município de Indianópolis – Minas Gerais

The objective of this study was to evaluate the occurrence of bovine mastitis by Staphylococcus sp., Streptococcus sp. and Candida sp. in a rural area of Indianopolis, Minas Gerais. It was realized the California Mastitis Test (CMT) in six collect, a total 671 of milk sample positive. Then the microbiological examination was performed, where the results revealed the presence of 137 milk samples with microbial multiplication. These, showed the presence of Staphylococcus aureus (45.2% of strains), other coagulase negative Staphylococcus (10.2%), Staphylococcus epidermidis (9.4%), Staphylococcus simulans (5.8%), other coagulase negative (15.3%), Streptococcus agalactiae (7.2%), other Streptococcus sp. (5.1%) and yeasts (1.4%). It was found that 100% of Staphylococcus were susceptible to rifampicin, gentamicin and ciprofloxacin; but, resistant to penicillin, tetracycline, and oxacillin. Regarding antimicrobial susceptibility Streptococcus, were employed, except to clindamycin, erythromycin and tetracycline. We conclude that there is a great necessity of proper hygiene practices and taking prophylactic measures taken in order to reduce the infection of animals caused by infectious microorganisms and resistance.

Minimum inhibitory concentrations of Staphylococcus aureus recovered from clinical and subclinical cases of bovine mastitis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)